Distribution, Persistence, and Efficacy of Adoptively Transferred Central and Effector Memory-Derived Autologous Simian Immunodeficiency Virus-Specific CD8+ T Cell Clones in Rhesus Macaques during Acute Infection

Abstract

Plasma viremia decreases coincident with the appearance of virus-specific CD8+ T cells during acute HIV or SIV infection. This finding, along with demonstrations of viral mutational escape from CD8+ T cell responses and transient increase in plasma viremia after depletion of CD8+ T cells in SIV-infected monkeys strongly suggest a role for CD8+ T cells in controlling HIV/SIV. However, direct quantitative or qualitative correlates between CD8+ T cell activity and virus control have not been established. To directly assess the impact of large numbers of virus-specific CD8+ T cells present at time of SIV infection, we transferred in vitro expanded autologous central and effector memory-derived Gag CM9-, Nef YY9-, and Vif WY8-specific CD8+ T cell clones to acutely infected rhesus macaques. The cells persisted in PBMCs between 4 and 9 d, but were not detected in gut-associated lymphoid tissue or lymph nodes. Interestingly, a high frequency of the infused cells localized to the lungs, where they persisted at high frequency for >6 wk. Although persisting cells in the lungs were Ag reactive, there was no measurable effect on virus load. Sequencing of virus from the animal receiving Nef YY9-specific CD8+ T cells demonstrated an escape mutation in this epitope <3 wk postinfection, consistent with immune selection pressure by the infused cells. These studies establish methods for adoptive transfer of autologous SIV-specific CD8+ T cells for evaluating immune control during acute infection and demonstrate that infused cells retain function and persist for at least 2 mo in specific tissues.

Adoptive Transfer of Engineered Rhesus Simian Immunodeficiency Virus-Specific CD8+T Cells Reduces the Number of Transmitted/Founder Viruses Established in Rhesus Macaques

“…T cells were cultured in RPMI 1640 media supplemented with 10% (vol/vol) fetal bovine serum (Gemini Bio Products), 10 mM HEPES buffer, 2 mM glutamine (Life Technologies), interleukin-2 (IL-2; NIH AIDS reagent repository) (50 IU/ml), penicillin (Life Technologies) (100 g/ml), and streptomycin (Life Technologies) (100 g/ml). Cells were expanded through biweekly stimulation with anti-CD3 (clone SP34-2; BD Biosciences) (30 ng/ml), IL-2 (50 IU/ml), irradiated human peripheral blood mononucleated cells (PBMC), and an Epstein-Barr virus-transformed B-cell line (TM B-LCL) (12,29,30).…”

“…Expanded transduced T cells from each animal were pooled and washed, and half the cells were labeled with 5 M CellTrace Violet (CTV; Life Technologies). Cells were resuspended in 50 ml saline solution supplemented with 2% autologous serum and infused into the femoral vein (2 ml/min) (12,21 The antibodies and tetramers used were as follows: CD3 (SP34-2), CD45 (HI30), CD4 (OKT4), CD8 (SK1), CCR7 (3D12), CD62L (SK11), CD28 (CD28.2), CD95 (DX2) (BD Biosciences), and ␣4-␤7 (11718; NIH) (34), NGFR; and ME20.4-1.H4 (Miltenyi Biotech), CM9 peptide MHC class I/tetramer (Beckman Coulter), and SL8 peptide/ MHC tetramer (NIH Tetramer Core Facility). Stimulated cells were stained for CD107a (H4A3) and for the intracellular cytokines gamma interferon (IFN-␥) (B27) and macrophage inflammatory protein 1 beta (MIP-1␤) (D21-1351; BD Biosciences).…”

“…Additionally, quantitative (11) or spatiotemporal (12) limitations could prevent effective antiviral CD8 ϩ T-cell responses from acting at foci of viral infection and replication, being too little, too late, and/or in the wrong place to affect the establishment of infection.…”

“…This powerful approach has been used less extensively to study the efficacy of anti-AIDS virus immune responses using SIV infection in a rhesus macaques model (12,(21)(22)(23)(24)(25)(26). A key challenge for these types of adoptive-transfer experiments is producing sufficient numbers of autologous cells for infusion.…”

Transplantation of mouse HSCs genetically modified to express a CD4-restricted TCR results in long-term immunity that destroys tumors and initiates spontaneous autoimmunity

“…Phenotypic changes in early to late effector T cells are associated with diminished in vivo effector function and persistence (3,4). Current strategies involving adoptive transfer of naive, central memory, or stem cell memory T cell populations have sought to take advantage of this continuum of functional change (5)(6)(7)(8). Despite these advances, however, it remains unclear what the stability and long-term immune reactivities of these populations will be, and how they can be translated to patient care.…”